FOLLOW-UP OF CASE OF ADVANCED SURVIVAL AND TRISOMY 18
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J. ment. Defic. Res. (1980) 24, 157 157 FOLLOW-UP OF CASE OF ADVANCED SURVIVAL AND TRISOMY 18 ARABELLA SMITH* and GESINA M. DEN DULK Cytogenetics Unit, Oliver Latham Laboratory, Health Commission of New South Wales, Australia INTRODUCTION In 1978 we reported in detail on an eleven-year-old girl with trisomy 18 (Smith, Silink and Ruxton, 1978). In the review of cases of advanced survival with trisomy 18 known at that time, there were three females still alive aged twenty-one, twenty- seven and six-and-a-half years. The girl reported by Stoll, Levy and Terrode (1974) is now aged eighteen (Stoll, 1978, personal communication). This review was only of cases considered to be non-mosaic. A further case of an eight-year-old girl has since been reported (Pauli, Pagon and Hall, 1978). Our patient was seen again at age twelve-and-a-half years, her physical para- meters have remained exactly as previously reported. As trisomy 18 had been ascer- tained on peripheral blood cultures, we obtained a skin culture on this occasion; the result of this culture showed 47,XX,+ 18. METHOD Tissue culture was performed according to a modification of the method described by Wolf (1974). A specimen of the skin was taken from the ventral surface of the forearm using local anaesthesia. In the laboratory, the specimen was bathed in chicken plasma and cut into small fragments (about 1 mm^). These were directly deposited on to the flat surface of a Lux ambitube. With the fiat surface of the explants rotated up, about 1 ml. of culture medium. Ham's FlO, hepes buffered, containing twenty per cent foetal calf serum, penicillin and streptomycin was added and each was gassed with carbon dioxide (5 per cent) in medical air. They were then incubated in this position at 37°C. After a few hours they were rotated back into normal position. After this they were left undisturbed for five days; every second day thereafter medium change and CO2 gassing was parformed. When the growth around the explant had become fairly confiuent and about 1 cm. in diameter, it was dispersed over the whole area by washing it twice with 0.2 per cent trypsin in calcium and magnesium-free Hanks balanced salt solution, with versene. It was left at room temperature for a few minutes before new culture medium was added. The cells were then dispersed by gently pipetting over the growth surface and reincubated. When subsequently growth had become overall confiuent, the above process was repeated, but a small amount of the culture medium *Reprint requests to Dr A. Smith, Cytogenetics Unit, Oliver Latham Laboratory, Health Commission of New South Wales, North Ryde, P.O. Box 53, New South Wales 2113, Australia. Received 13th February, 1980
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Transcript of FOLLOW-UP OF CASE OF ADVANCED SURVIVAL AND TRISOMY 18
J . ment. Defic. Res. (1980) 24, 157 157
FOLLOW-UP OF CASE OF ADVANCED SURVIVAL ANDTRISOMY 18
ARABELLA SMITH* and GESINA M. DEN DULK
Cytogenetics Unit, Oliver Latham Laboratory, Health Commission of New South Wales,Australia
INTRODUCTIONIn 1978 we reported in detail on an eleven-year-old girl with trisomy 18 (Smith,
Silink and Ruxton, 1978). In the review of cases of advanced survival with trisomy 18known at that time, there were three females still alive aged twenty-one, twenty-seven and six-and-a-half years. The girl reported by Stoll, Levy and Terrode (1974)is now aged eighteen (Stoll, 1978, personal communication). This review was onlyof cases considered to be non-mosaic. A further case of an eight-year-old girl hassince been reported (Pauli, Pagon and Hall, 1978).
Our patient was seen again at age twelve-and-a-half years, her physical para-meters have remained exactly as previously reported. As trisomy 18 had been ascer-tained on peripheral blood cultures, we obtained a skin culture on this occasion;the result of this culture showed 47,XX,+ 18.
METHODTissue culture was performed according to a modification of the method
described by Wolf (1974). A specimen of the skin was taken from the ventral surfaceof the forearm using local anaesthesia. In the laboratory, the specimen was bathed inchicken plasma and cut into small fragments (about 1 mm^). These were directlydeposited on to the flat surface of a Lux ambitube.
With the fiat surface of the explants rotated up, about 1 ml. of culture medium.Ham's FlO, hepes buffered, containing twenty per cent foetal calf serum, penicillinand streptomycin was added and each was gassed with carbon dioxide (5 per cent)in medical air. They were then incubated in this position at 37°C. After a few hoursthey were rotated back into normal position. After this they were left undisturbedfor five days; every second day thereafter medium change and CO2 gassing wasparformed.
When the growth around the explant had become fairly confiuent and about1 cm. in diameter, it was dispersed over the whole area by washing it twice with0.2 per cent trypsin in calcium and magnesium-free Hanks balanced salt solution,with versene. It was left at room temperature for a few minutes before new culturemedium was added. The cells were then dispersed by gently pipetting over thegrowth surface and reincubated. When subsequently growth had become overallconfiuent, the above process was repeated, but a small amount of the culture medium
*Reprint requests to Dr A. Smith, Cytogenetics Unit, Oliver Latham Laboratory,Health Commission of New South Wales, North Ryde, P.O. Box 53, New South Wales 2113,Australia.
Received 13th February, 1980
158 TRISOMY 18
with the dispersed cells was subcultured on coverslips in Leighton tubes for harvesting,making up the medium to 2 ml. per tube, mixing thoroughly and gassing. This wasallowed to grow for three days with a medium change on the second day. Colcemid(Gibco) to a final concentration of 0.5 ug/ml was added and allowed to act for fourhours. Hypotonic treatment with l-in-5 diluted medium for fifteen minutes followedand then fixation with Carnoys fixative. The processing was completed by stainingthe coverslips with Leishman and mounting inverted with DPX on to preparedslides.
DISCUSSIONCrippa, Marcoz, Klein and Bourquin (1978) considered that advanced survival
in cases of trisomy 18 was invariably associated with mosaicism. This was based ontheir finding of a small percentage of 46,XX normal cells in the skin culture of theirsixteen-and-a-half year old patient, whose peripheral blood culture showed only47,XX, +18 cells.
We wished to document the skin culture findings in our case. All cells examinedshowed 47,XX,+ 18 Karyotype. This finding does not exclude a normal cell line insome other circumscribed area of the body tissues (Goldstein, Kelly, Johanson andBlizzard, 1977). The somatic effect of such a small cell line on survival is unknown.
SUMMARYWe have reported the results of skin culture of a girl with trisomy 18, previously
reported when eleven years of age and now aged twelve-and-a-half years. There wasno evidence of mosaicism.
REFERENCESCRIPPA, L., MARCOZ, J. P., KLEIN, D. and BouRquiN, D. (1978 Les trisomies 18 a survie
prclongee (au-dela de 10 ans). Sont-elles, toutes des mosaiques. J. Genet. Hum. 26, 145.GOLDSTEIN, E. D., KELLY, E. T., JOHANSON, A. J. and BLIZZARD, R. M. (1977) Gonadal
dysgenesis with 45XO/46XX mosaicism demonstrated only in a streak gonad. J. Pediatr.90, 604.
PAULI, R. M., PAGON, R. A. and HALL, J. G. (1978) Trisomy 18 in sibs and maternal chromo-some 9 variant. Birth Defects. O.A.S. Vol. XIV, No. 6C, 297.
SMITH, A., SILINK, M. and RUXTON, J. (1978) Trisomy 18 in an 11-year-oId girl. J. ment.Defic. Res. 22, 277.
STOLL, C , LEVY, J. M. and TERRODE, E . (1974) Les survies prolongces dans la trisomie 18.Ann. Pediat. 21 , 185.
WOLF, U . (1974) Cell cultures from tissue explants. In: Methods in Human Cytogenetics. (Eds.)H. G. ScHWARz.\CHER and U. WOLF. Berlin, Heidelberg, New York: Springer Verlag,p. 45.
